In a letter to the editor of The New York Times, a top FBI official today disputed recent reports by ProPublica, PBS’ “Frontline” and McClatchy that challenged evidence in the agency’s investigation of the 2001 anthrax letter attacks and its conclusion that Army scientist Bruce Ivins was the perpetrator.

The letter from FBI Assistant Director for Public Affairs Michael P. Kortan followed editorials in The Times and The Washington Post that called for new, independent investigations into the anthrax case, which the FBI officially closed 18 months after Ivins committed suicide in the summer of 2008.

Kortan’s letter largely repeats prior claims in defense of the case, some of which were covered in our stories.

Here are key points from the letter and what we reported:

Ivins’ suspicious lab hours

Ivins was a top anthrax researcher at the U.S. Army Research Institute of Infectious Diseases in Fort Detrick, Md. He produced vaccines and tested them on lab animals. The FBI has said a spike in Ivins’ nighttime lab hours before the mailings explains when he prepared the letters.

What Kortan wrote: “[I]t was directly relevant that Dr. Ivins worked long hours alone during the time of the mailings in the laboratory’s ‘hot suites,’ where the anthrax that was genetically linked to the attack spores was produced and handled. He had not done that before the mailings, nor did he ever do it again.”

Ivins’ anthrax samples

Once the FBI determined the letters contained a strain of anthrax common in labs, the bureau asked scientists to submit samples of their spores. The FBI has said Ivins tried to hide his guilt by sending a false sample from his spore collection. Genetic tests later showed the spores in Ivins’ flask – RMR-1029 – matched the letter spores.

What we reported: An email shows that Ivins’ offered his spores for genetic tests in December 2001. Though samples he submitted from his flask in April 2002 were not a genetic match, Ivins maintained it was an innocent mistake. Furthermore, an FBI memo and Army document show that Ivins had given another sample of RMR-1029 to a different colleague around that time. Email messages and lab notebooks show that two years later Ivins helped a colleague collect a sample of RMR-1029 specifically for FBI evidence. Finally, the very first sample Ivins gave the FBI in February 2002 was rejected because it was in the wrong type of test tube. It was later tracked down, and tests determined it to be a close match to the letter spores.

What Kortan wrote: “Dr. Ivins submitted an intentionally misleading sample in April 2002 that was free of genetic markers. Samples of his anthrax spores that contained the genetic markers were either submitted before he realized the markers might trap him or were seized later by the F.B.I., not made available by Dr. Ivins.”

Was the anthrax treated?

FBI and other scientists who examined the attack spores found that they contained silicon, tin and some other elements. The presence of those elements fueled speculation that the spores were treated – weaponized – to make them more dispersible. The FBI has said its studies show the silicon was the result of a natural process.

What Kortan wrote: “[S]cientists directly involved in the lengthy investigation into the anthrax mailings — both from within the F.B.I. and outside experts — disagree with the notion that the chemicals in the mailed anthrax suggest more sophisticated manufacturing.”

Most news coverage of the National Academy of Sciences review of the FBI’s Amerithrax investigation has focused on its failure to conclusively show that Army scientist Bruce Ivins was the lone perpetrator of the 2001 anthrax attacks.
The academy report also detailed many significant methodological and organizational problems in the scientific portion of the Amerithrax probe.
If the FBI doesn’t address these scientific problems, its investigation of future bio-attacks may not support effective prosecutions or help a president decide how to respond, if the attack is state sponsored.

When the National Academy of Sciences issued its review of the FBI anthrax investigation earlier this year, the press fixated primarily on one point: The report found no conclusive evidence that Bruce Ivins, the Army scientist the government contends was responsible for a series of anthrax-laced letters mailed in 2001, produced them. Far less attention was devoted to the academy’s comments on the FBI’s problems with methodology — particularly the thoroughness of its research and its use of new technologies — and organization, especially at the start of the probe, when there was no structure for directing its scientific efforts. If the FBI fails to address these problems, it will remain ill-equipped to produce evidence for future bio-attack cases that stand up in court. Further, it will be unable to provide a president with sufficient information to support a possible retaliatory decision in the case of a state-sponsored bio-attack.

Perhaps the most astonishing result of the academy’s analysis involves the scope of the FBI’s findings in regard to the five anthrax letters at the heart of the case. After years of investigation, the bureau answered only a small number of the questions that it pursued: when the anthrax was produced, whether the material in all the letters was the same, and whether it had been weaponized. But other key questions relating to how and where the material was produced remained unanswered, making a determination of the material’s origin and the perpetrator’s identity problematic.

Early in the investigation, the FBI determined that the anthrax in the envelopes was of the virulent Ames strain and was produced in two separate batches with “differences in spore concentrations, color, contaminants, texture, growth media remnants and observed debris.” The presence of silicon in the material was initially deemed indicative of weaponization, as silicon dioxide has been used in past state bioweapons programs to enhance dispersion. Further analysis of the anthrax, however, indicated that the silicon was located within the spores, as the product of the spores’ natural growth process, rather than on their surface, where such dispersant material would normally reside. Carbon dating also determined that the spores were produced between 1998 and 2001, providing a possible chronological link with Ivins’s flask, labeled RMR-1029, which had been stored at US Army Medical Research Institute of Infectious Diseases (USAMRIID) since 1997. The FBI theorized that the flask w
as the source of the anthrax used in the 2001 attacks.

But genetic analysis was not able to ascertain the origin of the material in the five letters, which were sent to New York and Florida media outlets and to US senators Patrick Leahy and Thomas Daschle in Washington, DC. It also couldn’t identify the production methods used. Indeed, the academy’s analysis concluded that the RMR-1029 flask was not the immediate source of the letter material because it presented characteristics not shared with the letters. For example, unlike the anthrax spores in the letters, the RMR-1029 spores did not contain silicon in the coat. In addition, the RMR-1029 sample tested positive for the purification compounds meglumine and diatrizoate, which the letter samples did not contain, possibly indicating that a different purification method was used for the letter material. Therefore, the academy’s report concludes, if the letter material was indeed derived from the RMR-1029, several additional preparation steps involving cultivation, purification, an
d drying would have been necessary.

But the FBI investigation was not able to clarify what methods might have been used to grow, purify, concentrate and dry the anthrax sent in the letters. Given these uncertainties, the FBI was unable to determine the amount of time needed to produce the powdered anthrax, the type of skills and the equipment required to make it, the quality of the starting material, and the amount of liquid anthrax required to produce the five grams or so of dried anthrax in the letters.

Organizational issues. Probably the most important organizational flaw in the FBI probe was the lack of a coordinating structure at its outset. Because the FBI did not have the required scientific expertise to conduct an investigation of such breadth, 29 government, university, and private high-containment laboratories were commissioned to assist them. Several expert panels were also formed to review aspects of the scientific work, while additional outside expertise was sought from several government agencies. For example, during the first two years of the FBI investigation, the bureau did not create its own team to oversee and coordinate the work performed by all these actors, or to advise the FBI on the specific expertise required and the methodology choices available.

This changed in 2003, when the FBI created the Chemical, Biological, Radiological and Nuclear Sciences Unit, also known as CBSU. It seems, however, that the creation of CBSU did not solve all of the problems. For instance, the academy notes that near the end of its investigation, the FBI called on the expertise of statisticians; their involvement would have been useful throughout the process to provide advice on different scientific aspects of the case, from experimental design to data interpretation. Absent such assistance, and because it reached uncertain or ambiguous findings, the investigation naturally fell prey to controversy.

The 29 laboratories worked for the FBI in a compartmentalized fashion, unaware of what other labs were doing. This organizational choice was probably due to a dilemma the FBI faced: The scientists capable of assisting in the investigation could have also been potential suspects. Allowing them to review research conducted by other scientists could have tipped the perpetrator, or perpetrators, off. Still, compartmentalization runs counter to the scientific process, which relies on open exchange and collective interpretation of data.

Methodology issues. The academy’s report points to a number of methodology issues bearing on the thoroughness of the FBI’s research, its technical decision-making process, and its use of new technologies.

To help its investigation, the FBI created a repository of Ames anthrax strains. The purpose of the repository was to compare the anthrax in the letters to specimens obtained from various sources, in hopes of identifying the origin of the letter material. The FBI collected 1,070 specimens from 15 US organizations and three organizations in Great Britain, Canada, and Sweden, each having been requested to provide two samples of all the Ames strains they had in their collections. One sample was used for analysis, the other was archived. In addition, the US intelligence community provided samples from an unspecified foreign location (possibly Afghanistan, where Al Qaeda was suspected of developing anthrax).

The academy questioned how the FBI’s repository was established and populated. Because the Ames strain was widely used and exchanged among laboratories in the United States and overseas before 2001, the academy could not document that the FBI had identified all laboratories housing samples of the Ames strain. In addition, the academy found, the bureau did not provide the subpoenaed laboratories sufficiently precise instructions on how to prepare the samples or on the number of spores or cells required, and it did not supervise the preparation of the samples in these laboratories. Such methods suggested to the academy that the laboratories might not have used procedures consistent enough with one another to allow comparison, nor was there sufficient certainty that the subpoenaed laboratories produced samples of all the Ames strains in their possession.

The academy report also notes that it is not clear how the FBI made scientific decisions to pursue or not pursue some investigative routes or scientific approaches. For example, Lawrence Livermore National Laboratory’s analysis of the letter material revealed the presence of tin and iron that might have provided useful chemical signatures pointing to certain production methods. Yet, the academy found no evidence that this possibility was discussed or pursued by the FBI. Nor were alternative explanations for some findings apparently explored. For example, the FBI concluded the mutations found in the anthrax letters and Ivins’s RMR-1029 flask, pointed to a common origin. But the mutations could have been the result of parallel evolution rather than derivation, the national academy’s report says, and this possibility was not thoroughly investigated. The academy also raised concerns that material from some of the letters was not analyzed to determine whether it included the four
genetic markers used in comparing the samples.

Finally, the academy report notes that new technologies, such as “next generation” DNA sequencing, could have been more fully exploited to produce a more precise profile of the anthrax in the letters, strengthening the link with the RMR-1029 sample — or, perhaps, disproving it.

Recommendations for future investigations. Much of the academy’s report emphasizes the importance to biological investigations of a rigorous scientific approach — specifically a clearly defined decision-making process and well-defined methods. Methodology is particularly important; future attacks may involve agents that are not as well characterized as the Ames strain of anthrax, and the method used in the attack might not allow the recovery of a sufficient amount of agent for analysis. The academy report makes several recommendations to correct the organizational and methodological problems seen in the Amerithrax investigation.

As the academy report notes, “a scientific study is more than a series of well-executed experiments. The planning and decision-making used during the study are essential components of the science and can determine its outcome.” The report therefore recommends that in future investigations, an oversight organization should be created at the outset to “advise on procedures, strategies, and protocols, [and] help with new methods of scientific approaches.” This organization should be composed of experts not directly involved in the investigation and independent from the investigating organization, thereby fostering objective oversight and ongoing review of data results.

To ensure the thoroughness of future scientific investigations, the academy also suggests that relevant government agencies create and periodically update protocols and experimental designs for collecting and analyzing samples, preserving evidence, and otherwise buttressing the strength of biological cases in court. Equally important is the creation of a repository of all evidentiary material, so it can be compared with material in future events. The report emphasizes the importance of keeping abreast of new technological developments, as well; technological advances could not only support ongoing investigations, they could help in the review of past probes. For example, new genetic-sequencing technologies that emerged in the past decade could be used today to further investigate the Amerithrax samples, even though the criminal case is closed. Similar to the DNA analysis used so effectively in murder cases, these technologies could reinforce the case against Ivins, whose 2008
suicide precluded a public review of the case. Or the new technologies could point in a new direction altogether.

Finally, it is important to ask the question of what relative weight should be assigned to scientific information, as compared to other evidence gathered during a criminal investigation. The FBI indicated that questions relating to the production of the anthrax in the letters — whether someone had the skills required or the time needed for spore preparation — were not the only criteria used to eliminate suspects. But what weight was given to these criteria? Considering that the FBI never precisely established the required skills and spore preparation time, one may wonder whether the case would have been able to withstand the scrutiny of a trial.

To be sure, the field of microbial forensics is at an embryonic stage. If there is any overarching lesson to be taken from the FBI’s investigation, it is that microbial forensics, supported by rigorous adherence to sound methods and organizational discipline, is the sine qua non for determining the perpetrator of future bio-attacks. Indeed, allowing the perception that the United States cannot identify the source of an attack only increases the likelihood that a terrorist — or a country — might conduct such an attack. Deterrence of bio-attacks depends critically on absorbing this important lesson.

DXersaid

“Clearly confidential informants have their place in law enforcement, but only under the strictest control. The confidential informant agreement is not a license to commit crimes or to stand in the way of law enforcement. Turning a blind eye to murder to protect the flow of information makes those who do it accessories to the crime. The perceived worth of that policy of getting the bigger fish is just plain wrong. One can only hope the FBI has learned from Boston”

“Another valid concern is whether certain individuals should not be allowed to access select agents. While some groups have now been excluded categorically by the above-mentioned statutes, vigorous discussions concerning personnel reliability in a broader sense continue. Attention to an “insider threat” was amplified by the identification of Bruce Ivins, a scientist working in a U.S. government laboratory, as the possible perpetrator of the anthrax attacks.”

Comment: University of Michigan faced great controversy — that has not been made public — about the fact that they allowed a former Zawahiri associate to work alongside Bruce Ivins, Patricia Fellows (the scientist who won’t grant interviews who spun the passive mouse study the way she did) and their colleagues at the USAMRIID B3. He also worked with other anthrax (non-Ames) strains at LSU and conducted aerosol tests at Dugway using a simulant. The documents produced by the FBI show that Dr. Ivins did not know Dr. Hamouda was not a citizen when he came to work with them in the BL-3. The approval on the fly violated a long-standing memo on research by foreign nationals.

University of Michigan produced no documents in response to my request under FOIA. University of Michigan proved to be part of the problem and not part of the solution. Like GMU, it avoided the purposes of FOIA out of a self-interested financial self interest. (At GMU, a scientist coordinating with Anwar Awlaki shared a suite with the leading Ames researchers)

(In contrast, some university (in Montana perhaps?) that was involved in the SBIR for the Microbial Vac rushed a complete response.)

If you want to study the influence of money in sharp relief, then universities provide a clear case study.

The decision-makers at DOJ and FBI do not realize how many documents have already long ago gone out the door (and they won’t know until the whistleblowers come out of the woodwork). The IC has been focused since Dr. Ivins’ death on who screwed up Amerithrax and why?

The only safe harbor from the tsunami that will hit is to, as an individual, seek to produce all non-exempt documents under FOIA. You might just learn something and save your consequences of having withheld the documents earlier.

DXersaid

Under biographical information of the accomplished NAS panel members that reviewed the science in Amerithrax, you see Dr. Gast and Dr. Relman. Now we’ve heard from the next individual listed — this co-author and NAS panel member, Dr. Casadevall.

DXersaid

The two authors have previously written about the destruction of samples that occurred after the anthrax mailings. Of the two large collections destroyed, the scientists surveyed by the authors did not want to provide species identification of the destroyed samples.

In this study we have followed up on anecdotal and hearsay evidence that microbial collections were destroyed in the United
States following the imposition of the regulations associated with the Select Agents and Toxins List, to validate or refute that
information. Using a questionnaire, we documented 13 episodes of microbial collection destruction involving viral, bacterial,
and fungal strains, which we believe is almost certainly an underestimate of the number of collections destroyed. In every case,
the motivation for the destruction of the collection was a desire to avoid the perceived burdens of the regulatory environment
associated with operating under the Select Agent Regulations. Some institutions that destroyed isolates considered, and in
some cases tried, transferring their collections to registered institutions prior to collection destruction but desisted when
confronted with transport regulations. Destruction of microbial collections represents a loss of strains and biological diversity
available for biomedical research and future mechanistic, forensic, and epidemiologic investigations. Given the rapid evolution
of microbial strains, the destruction of archival collections is a potentially irretrievable loss that was an unintended
consequence of regulations to protect society against the nefarious use of biological agents. Furthermore, unregistered
institutions continue to destroy newly acquired clinical isolates, thus preventing the establishment of new repository collections.
We recommend that government agencies develop plans to ensure that microbial collections are preserved when
considering future additions to microbial threat lists under which the possession of certain microbes is criminalized.

***

Materials and Methods
The authors queried colleagues in the research community
with a letter that asked ‘‘for information as to whether such
microbial collections were destroyed in your institution in
the post 9/11 days as the select agent regulations’’ became
law. We asked 4 questions:
1. Are you aware whether microbial collections were destroyed
in your institution?
2. If yes, what types of samples were destroyed (pathogen
names would be helpful)?
3. Are you aware of whether there was an attempt to save
the collection by transferring it to a registered institution?
4. If a paper is published with this information, do you
have any problem with the identification of your institution
and/or your name as the information source? If
yes, please provide the above information, and we will
not name you or your institution in the report.
This letter was distributed to 1,000 individuals in the
biosafety community through an American Biological
Safety Association (ABSA) listserv, and to another group of
individuals through the Center for Science and Technology
Policy at AAAS.

***

Our goal was to obtain evidence that microbial collections
had indeed been destroyed in response to SATL regulations.
We received 13 affirmative responses of microbial collection
destruction in response to regulations associated with SATL
(Table 1). The geographic distribution of these affirmative
responses was as follows: Midwest 5, Northeast 3, South 1,
andWest 4. In addition, we received 5 responses stating that
no destruction had occurred in those institutions, and 4
responses stating that the investigators had successfully
managed to save their collections by moving them to SATLregistered
institutions.

We discerned considerable angst in the responses we
received, with most respondents asking for anonymity. We
suspect that concern about even participating in our survey
could have significantly reduced the number of respondents.
This anxiety ranged from asking for anonymity to
one institution’s refusing to disclose the identity of the
destroyed material and simply stating that it was bacteria.
Another individual provided the information in an anonymous
letter. We can only speculate as to the causes of
anxiety over responding to our questionnaire, but we note
that at least one correspondent worried about legalities, and
it is our impression that most contributors just wanted to
remain anonymous.

***

Two large collections destroyed. One involved archival material
from 1940-2002 and the other isolates from the 1950s to
the 1980s. Investigators did not want to provide species
identification for destroyed bacteria.

Arturo Casadevall, MD, PhD, is a Professor, Department of Medicine, Division of Infectious Diseases, and a Professor and the Leo and
Julia Forchheimer Chair in Microbiology & Immunology, Albert Einstein College of Medicine, Bronx, New York. Michael J. Imperiale,
PhD, is a Professor in the Department of Microbiology and Immunology, University ofMichiganMedical School, Ann Arbor, Michigan.

DXersaid

Condoleezza Rice’s book will be released on November 1, 2011. Perhaps some stores are less strict in observing the date of release and we could post any pertinent excerpts on anthrax sooner rather than later.